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The Journal of Cell Biology, Vol 61, 83-94, Copyright © 1974 by Rockefeller University Press

ARTICLE

ISOLATION AND CHARACTERIZATION OF LUMINAL MEMBRANES FROM URINARY BLADDER



J. Vergara 1, F. Zambrano 1, J. D. Robertson 1, and H. Elrod 1

1 From the Department of Anatomy, Duke University, Durham, North Carolina 27710 and the Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37203

A method is reported for the isolation of a highly purified fraction of urinary bladder membranes containing hexagonal plaques. The method uses zonal centrifugation as the final step of fractionation.

The purified fraction was characterized by its electron microscopic morphology, by its enzymatic profile, by quantitative and qualitative analysis of lipids and by the protein pattern obtained by electrophoresis in polyacrylamide sodium dodecyl sulfate gels. The fraction contains 65% lipids and 35% proteins. The major protein component has a molecular weight of 27,000 daltons.

Phospholipids are more than the 54% of the total lipid weight. Phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol are the major phospholipids with 50%, 30%, and 7% of the total lipid phosphorus, respectively.

The glycolipid fraction is 10% of the total lipid weight and is formed by only two components, both sulfatides. Total cholesterol makes up 36% of the total neutral lipid fraction of which cholesterol esters constitute 6%. Glycoproteins are also found to be present in the fraction.

Submitted on September 18, 1973
Revised on November 26, 1973


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